CN104458544B - A kind of liquid fluid system of flow cytometer - Google Patents

Abstract

A liquid flow system of a flow cytometer, relating to the field of flow cytometers, which solves the problem that the liquid flow system of the existing cytometer cannot guarantee the performance of the cytometer under the conditions of low sample injection rate and high sample injection rate. The problem of high detection accuracy. Includes two auxiliary sheath pumps, sample pump, bead pump, main sheath pump, wash pump, four solenoid valves, five pressure relief valves, five filters, six reversing valves, multiple tees, Flow Cell, Pulsation Buffer, Bubble Detection Module, Vent Module, Air Filter, Rinse Buffer, Sample Injection Line, Bead Injection Line, Waste Reservoir, Sheath Reservoir, Wash Reservoir, Disinfectant Reservoir, and Detox Reservoir Bubble pool. The invention ensures that the cytometer has high detection accuracy at both low sample injection rate and high sample injection rate, and has the functions of cleaning, disinfection and debubbling, which not only ensures the purity of different samples in the liquid flow system, In addition, the fluid stability and linearity of the sample liquid passing through the laser inspection area are improved.

Description

A liquid flow system of a flow cytometer

technical field

The invention relates to the technical field of flow cytometer, in particular to a liquid flow system of the flow cytometer.

Background technique

Flow cytometry can perform multi-parameter, quantitative analysis or sorting of cells or biological particles in a fast-flowing state. It has been widely used in basic research such as cell biology, immunology, physiology, and molecular biology. It is used in medical clinical diagnosis, environmental detection and so on. The liquid flow system is one of the three core systems of the flow cytometer. Its main function is to arrange the cells or other particles in the sample suspension in a single row and pass through the laser detection area sequentially and orderly. The liquid flow system mainly includes two parts, the flow chamber and the liquid path. The flow chamber uses the principle of hydrodynamic focusing to converge the sheath liquid and the sample liquid in the flow chamber, so that the sample liquid forms a stable laminar flow, commonly known as nuclear flow. At this time, the sheath liquid The liquid surrounds the sample liquid, and the sample liquid forms a stable linear flow liquid through the laser detection area for optical system detection and analysis. The main function of the liquid path is to ensure the accurate injection of the sheath fluid and the sample fluid. For the same liquid flow system, when the sample liquid injection rate increases, the diameter of the nuclear flow becomes larger, causing the cells or particles to be measured in the nuclear flow to easily deviate from the center of the irradiation spot, resulting in uneven irradiation energy, increased signal changes and The quality of the data is degraded, thereby reducing the detection accuracy of the instrument. Therefore, the high detection accuracy of the flow cytometer corresponds to the low sampling rate, and the low detection accuracy corresponds to the high sampling rate. If the user wants to obtain a high sample sampling rate and improve the detection efficiency, he must sacrifice the detection accuracy. A liquid flow system of a flow cytometer capable of maintaining high detection accuracy at both low and high sampling rates has very practical value.

Contents of the invention

In order to solve the problem that the existing cytometer liquid flow system cannot ensure that the cytometer has high detection accuracy under the conditions of low sample injection rate and high sample injection rate, the invention provides a flow cytometer liquid flow system. system.

The technical scheme that the present invention adopts for solving technical problems is as follows:

A liquid flow system of a flow cytometer of the present invention comprises a first auxiliary sheath fluid pump, a second auxiliary sheath fluid pump, a sample pump, a microbead pump, a main sheath fluid pump, a flushing pump, a first solenoid valve, a second Second solenoid valve, third solenoid valve, fourth solenoid valve, first pressure relief valve, second pressure relief valve, third pressure relief valve, fourth pressure relief valve, fifth pressure relief valve, first filter, first pressure relief valve The second filter, the third filter, the fourth filter, the fifth filter, the first reversing valve, the second reversing valve, the third reversing valve, the fourth reversing valve, the fifth reversing valve, the fifth reversing valve Six-way valve, multiple tees, flow cell, pulsation buffer, air bubble detection module, vent module, air filter, flush buffer, sample injection line, bead injection line, waste reservoir, sheath fluid reservoir , cleaning solution pool, disinfectant solution pool and degassing solution pool;

The flow chamber is composed of an inlet area, a converging area, an adjustment area, a detection area, and an auxiliary area that are closely connected in sequence. The upper end of the inlet area is provided with a main sheath fluid inlet, a sample inflow inlet, and an air pressure adjustment port. The adjustment area includes an upper and lower The integrated adjustment area whose ends are closely connected with the lower end of the convergence area and the upper end of the detection area, respectively, the entrance of the first adjustment area and the entrance of the second adjustment area are respectively closely connected with the left and right ends of the integrated adjustment area;

The flushing pump is respectively connected to the first interface of the flushing buffer and the sheath fluid pool through the liquid flow pipeline and the fifth reversing valve; the second interface of the flushing buffer is connected through the liquid flow pipeline, the fourth filter and the fourth The pressure relief valve is connected to the ventilation module, and the third interface is divided into two paths through a tee, one path is connected to the third interface of the air bubble detection module through the liquid flow pipeline and the fourth solenoid valve, and the other path is divided into two paths through the tee, and one path is passed through The liquid flow pipeline and the second electromagnetic valve are connected to the piston of the microbead pump, and the other is connected to the piston of the sample pump through the liquid flow pipeline and the third electromagnetic valve; the sample pump is connected to the piston of the sample pump through the liquid flow pipeline and the fourth reversing The valves are respectively connected to the sample injection tube and the sample inlet; the microbead pump is respectively connected to the microbead injection tube and the sample inlet through the liquid flow pipeline and the third reversing valve;

The main sheath liquid pump is divided into two paths through a tee, one path is connected to the ventilation module through the liquid flow pipeline, the third filter and the third pressure relief valve, and the other path is connected to the sixth reversing valve through the liquid flow pipeline. The sixth reversing valve is connected to the flushing buffer zone through the liquid flow pipeline and the fourth electromagnetic valve, and the sixth reversing valve is respectively connected to the third interface of the bubble detection module and the ventilation module through the liquid flow pipeline. , sheath liquid pool, cleaning liquid pool, disinfectant pool and degassing pool;

The first interface of the air bubble detection module is connected to the main sheath liquid inlet through the liquid flow pipeline and the pulsation buffer, and the second interface is connected to the ventilation module through the liquid flow pipeline and the first electromagnetic valve; The pipeline is connected to the air filter and the waste liquid tank, and a standard atmospheric pressure is maintained inside the ventilation module;

The first auxiliary sheath fluid pump is divided into two paths through a tee, one path is connected to the ventilation module through the liquid flow pipeline, the first filter and the first pressure relief valve, and the other path is connected to the ventilation module through the liquid flow pipeline and the first reversing valve respectively connected to the inlet of the first adjustment area of the flow chamber and the sheath liquid pool; the second auxiliary sheath liquid pump is divided into two paths through a tee, and one path is connected to the The ventilation module, the other way is connected to the inlet of the second adjustment area of the flow chamber and the sheath fluid pool respectively through the liquid flow pipeline and the second reversing valve; and a fifth pressure relief valve are connected to the vent module, and the auxiliary area of the flow chamber is connected to the waste liquid reservoir through the liquid flow line.

The interior of the entrance area is a cylindrical hollow structure; the interior of the convergence area is a conical hollow structure, and the diameters of the cross-sectional circles in the interior of the convergence area gradually decrease from top to bottom, and the maximum cross-sectional diameter is equal to the diameter of the internal cross-section circle of the entrance area. , the minimum cross-sectional circle diameter is 60-300 microns.

The internal structure of the entrance of the first adjustment area is the same as that of the entrance of the second adjustment area; the inside of the large diameter end of the entrance of the first adjustment area is a cylindrical hollow structure, and the inside of the small diameter end is a cylindrical hollow structure or a square hollow structure, The inside of the middle is a conical hollow structure, and the diameter of each cross-sectional circle inside the middle gradually decreases from the outside to the direction of the comprehensive adjustment area, and the maximum diameter of the cross-section circle is equal to the diameter of the inner cross-section circle of the large diameter end.

The detection area is made of quartz glass, the inside is a rectangular pipe or cylindrical pipe, and the outside is a cuboid or cylinder.

Both the inlet area and the converging area are made of resin or quartz glass.

The comprehensive adjustment area has the same shape and size as the internal section of the detection area, and the total length of the comprehensive adjustment area is smaller than that of the detection area.

The external shape of the auxiliary area is a cuboid, cube or cylinder; the lower end of the auxiliary area is a cylindrical hollow structure, the upper end is a conical hollow structure, and the diameter of each section circle inside the upper end gradually increases from top to bottom. The circle diameter is equal to the inner section circle diameter of the lower end.

The flushing pump adopts a syringe pump driven by a stepping motor with a volume of 600-1000 ml; both the sample pump and the bead pump adopt a syringe pump driven by a stepping motor with a volume of 500-600 ml. microliter; the first auxiliary sheath liquid pump, the second auxiliary sheath liquid pump and the main sheath liquid pump all use a stepping motor to drive a syringe pump with a syringe structure, with a volume of 30-40 ml.

The air bubble detection module is made of quartz glass or organic resin, and is a cuboid or cube structure with a square channel inside, and three interfaces are left on the outside to communicate with the internal channel; the ventilation module is a cuboid structure or a cube structure, with a square channel inside. There are multiple interfaces on the outside to connect to the internal channel;

The flushing buffer zone is made of quartz glass or organic resin, and has a cuboid or cube structure, with a square channel inside and three interfaces on the outside to communicate with the inner channel.

When the liquid flow system is in the low sample injection rate working mode, the first auxiliary sheath liquid pump and the second auxiliary sheath liquid pump do not work, the inlet of the first adjustment area and the inlet of the second adjustment area are closed, and the main sheath liquid pump passes through the sixth The reversing valve sucks the main sheath liquid from the sheath liquid pool, and the main sheath liquid pump pushes the main sheath liquid into the main sheath liquid inlet and enters the inlet area. The sample pump sucks the sample liquid from the sample tube through the fourth reversing valve, and the sample The pump pushes the sample liquid from the sample flow tube into the converging area. If microbeads need to be detected, the microbead pump sucks the microbead liquid from the microbead sampling tube through the third reversing valve, and the microbead pump pushes the microbead liquid from the sample The flow tube flows into the converging area, where the main sheath liquid, sample liquid and microbead liquid are converged by the principle of hydrodynamic focusing and form a stable laminar flow. After detection, it flows into the waste liquid pool from the auxiliary area; when the liquid flow system is in the high sample injection rate working mode, the inlet of the first adjustment area and the inlet of the second adjustment area are connected, and the first auxiliary sheath liquid pump and The second auxiliary sheath liquid pump sucks the auxiliary sheath liquid from the sheath liquid pool through the first reversing valve and the second reversing valve respectively, and the auxiliary sheath liquid enters the comprehensive adjustment area from the inlet of the first adjustment area and the inlet of the second adjustment area respectively , keep the flow rate of main sheath liquid and sample liquid constant, adjust the flow rate of auxiliary sheath liquid to compress the stable laminar flow flowing in from the converging area, the sample liquid maintains stable laminar flow after being compressed, and the compressed stable laminar flow enters To the detection area, after detection, it flows into the waste liquid pool from the auxiliary area, the flow rate of the main sheath liquid is greater than or equal to the flow rate of the auxiliary sheath liquid, and the flow rate of the auxiliary sheath liquid is greater than the sampling rate of the sample liquid.

The beneficial effects of the present invention are:

The liquid flow system of the present invention has the functions of cleaning, disinfection, and debubbling, which not only ensures the purity of different samples in the liquid flow system, but also improves the fluid stability and linearity of the sample liquid when it passes through the laser inspection area, which is beneficial to Improve the detection accuracy and instrument stability of flow cytometer. In addition, the present invention has two working modes of low speed and high speed, which not only ensures high detection accuracy at both low sample injection rate and high sample injection rate, but also realizes the reduction of sample liquid at high sample injection rate. The stable laminar flow diameter after convergence ensures that the instrument has high detection accuracy.

Description of drawings

FIG. 1 is a schematic structural diagram of a liquid flow system of a flow cytometer of the present invention.

Fig. 2 is a schematic diagram of the structure of the flow chamber.

In the figure: 1. Inlet area, 2. Convergence area, 3. Adjustment area, 4. Detection area, 5. Auxiliary area, 6. Main sheath liquid inlet, 7. Sample flow inlet, 8. Air pressure adjustment port, 9. Second Inlet of the first adjustment area, 10, entrance of the second adjustment area, 11, comprehensive adjustment area, 12, sample flow tube, 15, first reversing valve, 16, second reversing valve, 17, first auxiliary sheath liquid pump, 18. Second auxiliary sheath liquid pump, 19. First filter, 20. First pressure relief valve, 21. Second filter, 22. Second pressure relief valve, 23. Pulsation buffer, 24. Bubble detection module , 25, the first solenoid valve, 26, the ventilation module, 27, the air filter, 28, the sample pump, 29, the bead pump, 30, the main sheath liquid pump, 31, the washing pump, 32, the second solenoid valve, 33 , the third solenoid valve, 34, the fourth solenoid valve, 35, the flushing buffer zone, 36, the third pressure relief valve, 37, the third filter, 38, the fourth pressure relief valve, 39, the fourth filter, 40 , the fifth filter, 41, the fifth pressure relief valve, 42, the third reversing valve, 43, the fourth reversing valve, 44, the fifth reversing valve, 45, the sixth reversing valve, 46, three-way , 47, sample injection tube, 48, microbead injection tube, 49, waste liquid pool, 50, sheath liquid pool, 51, cleaning liquid pool, 52, disinfectant liquid pool, 53, degassing liquid pool.

detailed description

The present invention will be described in further detail below in conjunction with the accompanying drawings.

As shown in Figure 1, the liquid flow system of a flow cytometer of the present invention includes a flow chamber, a fluid pump, a liquid flow pipeline, a solenoid valve, a pressure relief valve, a reversing valve, a pulsation buffer 23, and a bubble detection module 24. Ventilation module 26, air filter 27, flush buffer 35, sample injection tube 47, bead injection tube 48, waste liquid pool 49, sheath fluid pool 50, cleaning solution pool 51, disinfectant pool 52 and Bubble liquid pool 53.

As shown in Figure 2, the flow chamber is composed of an inlet area 1, a converging area 2, an adjustment area 3, a detection area 4, and an auxiliary area 5, and the inlet area 1, the convergence area 2, the adjustment area 3, the detection area 4, and the auxiliary area 5 tightly connected into one. The upper end of the inlet area 1 is provided with a main sheath liquid inlet 6, a sample inflow port 7 and an air pressure adjustment port 8, the sample inflow port 7 is located between the main sheath liquid inlet 6 and the air pressure adjustment port 8, and the sample inflow port 7 is used to place the sample flow tube 12. The sample flow tube 12 enters the inlet area 1 along the sample inflow port 7 and reaches the converging area 2. The air pressure adjustment port 8 is used to adjust the air pressure change in the flow chamber.

The interior of the entrance area 1 is a cylindrical hollow structure with a certain length. The internal section of the entrance area 1 is circular, and the diameters of the cross-sectional circles inside the entrance area 1 are the same; the external shape of the entrance area 1 is cuboid, cube or cylinder, etc., and the entrance area 1 There is a mechanical interface on the outside for installation and fixing. The entrance area 1 and the convergence area 2 are integrally formed by laser, the lower end of the entrance area 1 is in close contact with the upper end of the convergence area 2, and the materials of the entrance area 1 and the convergence area 2 are resin or quartz glass.

The inside of the converging area 2 is a conical hollow structure with a certain length. The internal section of the converging area 2 is circular, and the diameter of each cross-sectional circle inside the converging area 2 gradually decreases from top to bottom. 1. The diameter of the internal cross-sectional circle is the same. The minimum diameter of the internal cross-sectional circle of the converging area 2 is 60 to 300 microns. The specific size of the internal cross-sectional circle diameter of the converging area 2 is determined according to the size of the cells or particles to be tested; the external shape of the converging area 2 is cuboid, cube, Cylinder or cone, etc., the outer shape of the converging area 2 is preferably the same as that of the entrance area 1, which is beneficial to the laser integrated molding process, or it can be different, and it can be designed according to the needs.

The adjustment area 3 is provided with three parts: the first adjustment area entrance 9, the second adjustment area entrance 10 and the comprehensive adjustment area 11. The external shapes of the first adjustment area entrance 9 and the second adjustment area entrance 10 are cuboid, cube, cylinder or Cone, etc., the first adjustment area entrance 9 and the second adjustment area entrance 10 external shapes can be the same or different; the first adjustment area entrance 9 internal structure is the same as the second adjustment area entrance 10 internal structure, the first adjustment area entrance 9. The front end is a large-diameter end. The inside of the front end of the entrance of the first adjustment area 9 is a cylindrical hollow structure with a certain length. It has a conical hollow structure with a certain length, the middle internal section is circular, and the diameter of each section circle in the middle decreases gradually from the outside to the comprehensive adjustment area 11. The diameter is the same; the rear end of the entrance 9 of the first adjustment area is a small-diameter end, and the interior of the rear end of the entrance 9 of the first adjustment area is a cylindrical hollow structure or a square hollow structure with a certain length; the upper end of the comprehensive adjustment area 11 passes through the transition area and the convergence area 2. The lower end is closely connected. The left end of the comprehensive adjustment area 11 is in close contact with the small-caliber end of the entrance 9 of the first adjustment area. Laser integral molding processing.

The outer shape of the detection area 4 is a cuboid or cylinder, etc. The material of the detection area 4 is quartz glass. The interior of the detection area 4 is a rectangular pipe or cylindrical pipe with a certain length. The internal cross section of the detection area 4 is circular or rectangular. Comprehensive adjustment The internal cross-sectional shape and size of area 11 are the same as the internal cross-sectional shape and size of detection area 4, and the overall length of comprehensive adjustment area 11 is shorter than detection area 4. When both detection area 4 and comprehensive adjustment area 11 are rectangular pipes with a certain length inside, the detection The internal sections of the area 4 and the comprehensive adjustment area 11 are both rectangular, and the length × width of the internal section of the comprehensive adjustment area 11 is equal to the length × width of the internal section of the detection area 4. When both the detection area 4 and the comprehensive adjustment area 11 have For a cylindrical pipeline of a certain length, the internal cross-sections of the detection zone 4 and the comprehensive adjustment zone 11 are both circular, and the internal cross-sectional diameter of the comprehensive adjustment zone 11 is equal to the internal cross-sectional diameter of the detection zone 4 .

The external shape of the auxiliary area 5 is a cuboid, a cube or a cylinder, etc., the upper end of the auxiliary area 5 is a small-caliber end, and the inner part of the upper end of the auxiliary area 5 is a conical hollow structure with a certain length, and the inner section of the upper end of the auxiliary area 5 is circular; The inside of the lower end of 5 is a cylindrical hollow structure with a certain length, the inner section of the lower end of the auxiliary area 5 is circular, and the diameters of the cross-sectional circles inside the lower end of the auxiliary area 5 are the same; the diameters of the cross-sectional circles inside the upper end of the auxiliary area 5 gradually increase from top to bottom Large, the maximum diameter of the internal cross-sectional circle at the upper end of the auxiliary area 5 is the same as the diameter of the inner cross-sectional circle at the lower end of the auxiliary area 5;

As shown in FIG. 1 , the ventilation module 26 is a cuboid or cube structure, with a square channel inside and a plurality of interfaces on the outside to communicate with the inner channel. The upper end interface of the ventilation module 26 is connected to the air filter 27 through the liquid flow pipeline to communicate with the atmosphere, and the lower end interface is connected to the waste liquid pool 49 through the liquid flow pipeline. The interior of the ventilation module 26 maintains a standard atmospheric pressure.

The fluid pump includes: a first auxiliary sheath liquid pump 17, a second auxiliary sheath liquid pump 18, a sample pump 28, a microbead pump 29, a main sheath liquid pump 30 and a flushing pump 31; the solenoid valve includes: a first solenoid valve 25, a second Two solenoid valves 32, the third solenoid valve 33, the fourth solenoid valve 34; the relief valves include: the first relief valve 20, the second relief valve 22, the third relief valve 36, the fourth relief valve 38, The fifth pressure relief valve 41; the filter includes: the first filter 19, the second filter 21, the third filter 37, the fourth filter 39, the fifth filter 40; the reversing valve includes: the first reversing Valve 15, the second reversing valve 16, the third reversing valve 42, the fourth reversing valve 43, the fifth reversing valve 44, the sixth reversing valve 45, wherein the first reversing valve 15, the second reversing valve The reversing valve 16, the third reversing valve 42, the fourth reversing valve 43 and the fifth reversing valve 44 are all four-position reversing valves, and the sixth reversing valve 45 is a six-position reversing valve.

As shown in Figure 1, the flushing pump 31 is connected to the first position of the fifth reversing valve 44 through the liquid flow pipeline, and the second position of the fifth reversing valve 44 is connected to the flushing buffer zone 35 through the liquid flow pipeline. The pipeline and the fifth reversing valve 44 are connected to the flushing buffer zone 35, the fourth position of the fifth reversing valve 44 is connected to the sheath fluid pool 50 through the liquid flow pipeline, and the third position of the fifth reversing valve 44 is empty. The flushing pump 31 is a syringe pump driven by a stepping motor with a volume of 600-1000 milliliters.

The flushing buffer zone 35 is a cuboid or cube structure with a square channel inside and three interfaces connected to the inner channel on the outside, and is made of quartz glass or organic resin. The first interface among the three interfaces of the flushing buffer zone 35 is connected to the second position of the fifth reversing valve 44 through the liquid flow pipeline, and the second interface is connected to the second position through the liquid flow pipeline, the fourth filter 39 and the fourth pressure relief valve 38 is connected to the ventilation module 26, and the third interface is divided into two paths through the tee 46, and one path is connected to the air bubble detection module 24 and the sixth solenoid valve 45 through the liquid flow pipeline and the fourth solenoid valve 34; The channel 46 is further divided into two paths, one path is connected to the piston of the microbead pump 29 through the liquid flow pipeline and the second electromagnetic valve 32, and the other path is connected to the piston of the sample pump 28 through the liquid flow pipeline and the third electromagnetic valve 33. The other end of the sample pump 28 is connected to the second position of the fourth reversing valve 43 through the liquid flow pipeline, and one position of the fourth reversing valve 43 is connected to the sample injection tube 47 through the liquid flow pipeline, and the third position of the fourth reversing valve 43 is Connect to the sample inflow port 7 of the flow cell through a fluid line. The sample pump 28 is a syringe pump driven by a stepping motor with a volume of 500-600 microliters. The other end of the microbead pump 29 is connected to the second position of the third reversing valve 42 through the liquid flow pipeline, and one position of the third reversing valve 42 is connected to the microbead sampling tube 48 through the liquid flow pipeline, and the third reversing valve 42 The three positions are connected to the sample inflow port 7 of the flow cell by fluid lines. The microbead pump 29 is a syringe pump driven by a stepping motor with a volume of 500-600 microliters.

The main sheath liquid pump 30 is divided into two circuits through the tee 46, one circuit is connected to the ventilation module 26 through the liquid flow pipeline, the third filter 37 and the third pressure relief valve 36, and the other circuit is connected to the sixth valve through the liquid flow pipeline. The long-pass end of the valve 45, the sixth reversing valve 45 is connected to the sheath fluid pool 50 through the liquid flow pipeline, and the second position of the sixth reversing valve 45 is connected to the bubble detection module 24 and the flushing buffer zone through the liquid flow pipeline 35. The third position of the sixth reversing valve 45 is connected to the ventilation module 26 through the liquid flow pipeline, and the fourth, fifth and sixth positions of the sixth reversing valve 45 are respectively connected to the cleaning liquid pool 51 and the degassing Liquid pond 53 and disinfectant liquid pond 52. The main sheath fluid pump 30 is a syringe pump driven by a stepping motor with a volume of 30-40 ml.

The air bubble detection module 24 is a cuboid or cube structure, with a square channel inside and three interfaces on the outside to communicate with the inner channel, and is made of quartz glass or organic resin. The first interface among the three interfaces of the air bubble detection module 24 is connected to the main sheath liquid inlet 6 of the flow chamber through the liquid flow pipeline and the pulsation buffer 23, and the second interface is connected to the first solenoid valve 25 through the liquid flow pipeline To the vent module 26 , the third port is connected to the flushing buffer zone 35 through the liquid flow line and the fourth solenoid valve 34 .

The first auxiliary sheath liquid pump 17 is divided into two paths through the tee 46, one path is connected to the ventilation module 26 through the liquid flow pipeline, the first filter 19 and the first pressure relief valve 20, and the other path is connected to the first through the liquid flow pipeline. One reversing valve 15, one position, the second position of the first reversing valve 15 is connected to the first adjustment area inlet 9 of the flow chamber through the liquid flow pipeline, and the three positions of the first reversing valve 15 are connected to the sheath fluid through the liquid flow pipeline Pool 50, the four positions of the first reversing valve 15 are vacant. The first auxiliary sheath fluid pump 17 is a syringe pump driven by a stepping motor with a volume of 30-40 ml.

The second auxiliary sheath liquid pump 18 is divided into two paths through the tee 46, one path is connected to the ventilation module 26 through the liquid flow pipeline, the second filter 21 and the second pressure relief valve 22, and the other path is connected to the first through the liquid flow pipeline. The second reversing valve 16 is one, the second reversing valve 16 is connected to the second adjustment area inlet 10 of the flow chamber through the liquid flow pipeline, and the second reversing valve 16 is connected to the sheath liquid through the liquid flow pipeline. Pool 50, the second reversing valve 16 four positions are vacant. The second auxiliary sheath fluid pump 18 is a syringe pump driven by a stepping motor with a volume of 30-40 ml.

The air pressure regulating port 8 of the flow chamber is connected to the ventilation module 26 through the liquid flow pipeline, the fifth filter 40 and the fifth pressure relief valve 41 , and the outlet of the auxiliary area 5 of the flow chamber is connected to the waste liquid pool 49 through the liquid flow pipeline.

During detection, the fourth reversing valve 43 is controlled so that the sample pump 28 is connected to the sample injection tube 47, the sample pump 28 is started, and the sample liquid is sucked into the sample pump 28 from the sample injection tube 47; The valve 42 connects the microbead pump 29 with the microbead sampling pipe 48, and starts the microbead pump 29, and the microbead liquid is sucked into the microbead pump 29 from the microbead sampling pipe 48; the sixth reversing valve is controlled 45 Make the main sheath liquid pump 30 conduct with the sheath liquid pool 50, and start the main sheath liquid pump 30, the main sheath liquid is sucked into the main sheath liquid pump 30 from the sheath liquid pool 50; control the fourth reversing valve 43 to make The sample pump 28 is connected to the sample inlet 7, the third reversing valve 42 is controlled to make the microbead pump 29 connected to the sample inlet 7, and the sixth reversing valve 45 is controlled so that the main sheath liquid pump 30 is connected to the main sheath liquid inlet 6. conduction, control the sample pump 28 and the microbead pump 29 to push the sample liquid and the microbead liquid into the sample flow tube 12 located in the sample inflow port 7, and control the main sheath liquid pump 30 to push the main sheath liquid into the flow chamber, the main sheath liquid flows into the inlet area 1 from the main sheath liquid inlet 6, the sample flow tube 12 enters the inlet area 1 along the sample flow inlet 7 and reaches the converging area 2, the sample liquid and microbead liquid in the sample flow tube 12 flow into In converging area 2, the main sheath liquid, sample liquid and microbead liquid converge in converging area 2 through the principle of hydrodynamic focusing and form a stable laminar flow. At this time, the main sheath liquid flows around the sample liquid and microbead liquid, and then enters the Go to the comprehensive adjustment area 11, then enter the detection area 4 for detection, enter the auxiliary area 5 after detection, and flow into the waste liquid pool 49 from the outlet of the auxiliary area 5.

During cleaning, control the fifth reversing valve 44 to connect the flushing pump 31 to the sheath fluid pool 50, start the flushing pump 31 to suck flushing fluid from the sheath fluid pool 50, and control the fifth reversing valve 44 to connect the flushing pump 31 to the flushing buffer. The area 35 is turned on, and the flushing liquid enters the flushing buffer 35, and from the flushing buffer 35 respectively enters the sample pump 28, the microbead pump 29 and the flow chamber, and finally enters the waste liquid pool 49 to complete the cleaning function.

During disinfection, control the sixth reversing valve 45 to make the main sheath liquid pump 30 communicate with the disinfectant solution pool 52, start the main sheath liquid pump 30 to inhale disinfectant from the disinfectant liquid pool 52, and control the sixth reversing valve 45 to make the main sheath liquid The liquid pump 30 is connected to the flushing buffer 35, and the disinfectant enters the flushing buffer 35, and from the flushing buffer 35 enters the sample pump 28, the bead pump 29 and the flow chamber, and finally enters the waste liquid pool 49 to complete the disinfection function.

When removing air bubbles, open the first electromagnetic valve 25 to allow air to enter the flow chamber through the air filter 27 and the ventilation module 26, then close the first electromagnetic valve 25, and repeat the cleaning steps to discharge the air.

Adjust the conduction relationship of the first reversing valve 15, the second reversing valve 16, the third reversing valve 42, the fourth reversing valve 43, the fifth reversing valve 44, the sixth reversing valve 45 and the first auxiliary valve. Sheath liquid pump 17, the second auxiliary sheath liquid pump 18, sample pump 28, microbead pump 29, main sheath liquid pump 30 and flushing pump 31 working state, the liquid flow system of a kind of flow cytometer of the present invention has cleaning , disinfection, and de-bubble functions, which not only ensure the purity of different sample liquids in the liquid flow system, but also improve the fluid stability and linearity of the sample liquid when it passes through the laser inspection area, which is conducive to improving the detection of flow cytometers. precision and stability.

The liquid flow system of a flow cytometer of the present invention has at least two working modes, two working modes are cited in this embodiment, namely low-speed and high-speed working modes. When the liquid flow system is in the low sample injection rate working mode, the first auxiliary sheath liquid pump 17 and the second auxiliary sheath liquid pump 18 do not work, the first adjustment area inlet 9 and the second adjustment area inlet 10 are closed, and the main sheath liquid The pump 30 sucks the main sheath liquid from the sheath liquid pool 50 through the sixth reversing valve 45, and then the main sheath liquid is pushed by the main sheath liquid pump 30 to enter the main sheath liquid inlet 6 and enter the inlet area 1, and the sample pump 28 passes through the fourth changeover valve. The valve 43 sucks the sample liquid from the sample pipe 47, and then the sample pump 28 pushes the sample liquid to flow from the sample flow pipe 12 into the converging area 2. At this time, if microbeads need to be detected, the microbead pump 29 passes through the third reversing valve. 42 suck the microbead liquid from the microbead sampling tube 48, then push the microbead liquid from the sample flow tube 12 into the converging area 2 by the microbead pump 29, and the main sheath liquid, the sample liquid and the microbead liquid pass through in the converging area 2 The principle of hydrodynamic focusing converges and forms a stable laminar flow. At this time, the main sheath liquid flows around the sample liquid and microbead liquid, and then enters the comprehensive adjustment area 11, and then enters the detection area 4 for detection, and then enters the auxiliary sheath liquid. Zone 5 and flows into the waste liquid pool 49 from the outlet of the auxiliary zone 5; when the liquid flow system is in the high sample injection rate working mode, the first adjustment zone inlet 9 and the second adjustment zone inlet 10 are connected, and the first auxiliary sheath liquid pump 17 and the second auxiliary sheath liquid pump 18 suck auxiliary sheath liquid from the sheath liquid pool 50 through the first reversing valve 15 and the second reversing valve 16 respectively, and the auxiliary sheath liquid is drawn from the inlet 9 of the first adjustment area and the second adjustment area The inlets 10 enter the comprehensive adjustment area 11 respectively, keep the flow rate of the main sheath liquid and the sample liquid constant, adjust the flow rate of the auxiliary sheath liquid to compress the stable laminar flow flowing from the converging area 2, and maintain a stable layer after the sample liquid is compressed flow, but the diameter becomes smaller, the cell movement speed is faster, the compressed stable laminar flow enters the detection area 4, and finally flows into the waste liquid pool 49 from the auxiliary area 5. The flow rate of the auxiliary sheath liquid is greater than the sampling rate of the sample liquid, and the flow rate of the main sheath liquid is greater than or equal to the flow rate of the auxiliary sheath liquid.

The liquid flow system of a flow cytometer of the present invention adopts at least two working modes, which not only ensures the high detection accuracy of low sample injection rate, but also realizes the reduction of sample liquid after converging at high sample injection rate. The stable laminar flow diameter ensures that the instrument has high detection accuracy.

Claims (10)

1. a liquid flow system of flow cytometer, it is characterized in that, comprises the first auxiliary sheath liquid pump (17), the second auxiliary sheath liquid pump (18), sample pump (28), microbead pump (29) , main sheath fluid pump (30), flushing pump (31), first solenoid valve (25), second solenoid valve (32), third solenoid valve (33), fourth solenoid valve (34), first discharge valve Pressure valve (20), second pressure relief valve (22), third pressure relief valve (36), fourth pressure relief valve (38), fifth pressure relief valve (41), first filter (19), The second filter (21), the third filter (37), the fourth filter (39), the fifth filter (40), the first reversing valve (15), the second reversing valve (16), Third reversing valve (42), fourth reversing valve (43), fifth reversing valve (44), sixth reversing valve (45), multiple tees (46), flow chamber, pulsation damper (23), air bubble detection module (24), ventilation module (26), air filter (27), flush buffer (35), sample injection tube (47), microbead injection tube (48), waste liquid Pool (49), sheath fluid pool (50), cleaning fluid pool (51), disinfectant pool (52) and degassing pool (53); The flow chamber is composed of an inlet area (1), a converging area (2), an adjustment area (3), a detection area (4) and an auxiliary area (5) which are closely connected in sequence. The upper end of the inlet area (1) is provided with a main The sheath liquid inlet (6), the sample inlet (7) and the air pressure adjustment port (8), the adjustment area (3) includes a comprehensive adjustment in which the upper and lower ends are respectively in close contact with the lower end of the converging area (2) and the upper end of the detection area (4) zone (11), the entrance of the first adjustment zone (9) and the entrance of the second adjustment zone (10) which are closely connected with the left and right ends of the comprehensive adjustment zone (11); The flushing pump (31) is respectively connected to the first interface of the flushing buffer (35) and the sheath fluid pool (50) through the liquid flow pipeline and the fifth reversing valve (44); the first interface of the flushing buffer (35) The second interface is connected to the ventilation module (26) through the liquid flow pipeline, the fourth filter (39) and the fourth pressure relief valve (38). The fourth solenoid valve (34) is connected to the third interface of the air bubble detection module (24), and the other path is divided into two paths through the tee two, and one path is connected to the microbead pump ( 29), the other way is connected to the piston of the sample pump (28) through the liquid flow pipeline and the third electromagnetic valve (33); ) are respectively connected to the sample injection tube (47) and the sample inlet (7); the microbead pump (29) is connected to the microbead injection tube ( 48) and sample inflow port (7); The main sheath liquid pump (30) is divided into two paths through a three-way connection, one path is connected to the ventilation module (26) through a liquid flow pipeline, a third filter (37) and a third pressure relief valve (36), and the other path Connect to the long-pass end of the sixth reversing valve (45) through the liquid flow pipeline, and the sixth reversing valve (45) is connected to the flushing buffer zone (35) through the liquid flow pipeline and the fourth solenoid valve (34) , the sixth reversing valve (45) is respectively connected to the third interface of the air bubble detection module (24), the ventilation module (26), the sheath liquid pool (50), the cleaning liquid pool (51), the disinfection Liquid pool (52) and degassing liquid pool (53); The first interface of the air bubble detection module (24) is connected to the main sheath liquid inlet (6) through the liquid flow pipeline and the pulsation buffer (23), and the second interface is connected to the first electromagnetic valve (25) through the liquid flow pipeline To the ventilation module (26); the ventilation module (26) is connected to the air filter (27) and the waste liquid pool (49) through the liquid flow pipeline respectively, and a standard atmospheric pressure is maintained inside the ventilation module (26); The first auxiliary sheath liquid pump (17) is divided into two paths through a three-way connection, and one path is connected to the ventilation module (26) through a liquid flow pipeline, a first filter (19) and a first pressure relief valve (20), The other way is respectively connected to the first adjustment area inlet (9) and the sheath liquid pool (50) of the flow chamber through the liquid flow pipeline and the first reversing valve (15); the second auxiliary sheath liquid pump (18) passes through The three-way is divided into two ways, one way is connected to the ventilation module (26) through the liquid flow pipeline, the second filter (21) and the second pressure relief valve (22), and the other way is connected to the ventilation module (26) through the liquid flow pipeline and the second reversing The valve (16) is respectively connected to the second adjustment area inlet (10) of the flow chamber and the sheath fluid pool (50); the air pressure adjustment port (8) of the flow chamber passes through the liquid flow pipeline, the fifth filter (40) and a fifth pressure relief valve (41) are connected to the vent module (26), and the auxiliary area (5) of the flow chamber is connected to the waste liquid reservoir (49) through a liquid flow line. 2. The liquid flow system of a flow cytometer according to claim 1, characterized in that, the inside of the inlet area (1) is a cylindrical hollow structure; the inside of the converging area (2) is a conical hollow structure, the diameter of each cross-sectional circle inside the converging area (2) gradually decreases from top to bottom, the largest cross-sectional circle diameter is equal to the internal cross-sectional circle diameter of the entrance area (1), and the smallest cross-sectional circle diameter is 60-300 microns. 3. The liquid flow system of a kind of flow cytometer according to claim 1, characterized in that, the internal structure of the entrance of the first adjustment area (9) is the same as that of the entrance of the second adjustment area (10); The entrance of the first adjustment zone (9) has a cylindrical hollow structure inside the large-diameter end, a cylindrical hollow structure or a square hollow structure inside the small-diameter end, and a conical hollow structure inside the middle, and the diameters of the cross-sectional circles in the middle are integrated from the outside to the outside. The direction of the adjustment area (11) decreases gradually, and the maximum cross-sectional circle diameter is equal to the internal cross-sectional circle diameter of the large-diameter end. 4. The liquid flow system of a flow cytometer according to claim 1, wherein the detection area (4) is made of quartz glass, the inside is a rectangular pipe or a cylindrical pipe, and the outside is a cuboid or cylinder. 5. The liquid flow system of a flow cytometer according to claim 1, characterized in that, both the inlet area (1) and the converging area (2) are made of resin or quartz glass. 6. The liquid flow system of a kind of flow cytometer according to claim 1, characterized in that, the shape and size of the internal section of the comprehensive adjustment area (11) and the detection area (4) are the same, and the comprehensive adjustment The total length of the zone (11) is smaller than that of the detection zone (4). 7. The liquid flow system of a flow cytometer according to claim 1, characterized in that, the external shape of the auxiliary area (5) is a cuboid, cube or cylinder; the inner part of the lower end of the auxiliary area (5) is It is a cylindrical hollow structure, and the inside of the upper end is a conical hollow structure. The diameter of each cross-sectional circle inside the upper end gradually increases from top to bottom, and the maximum cross-sectional circle diameter is equal to the inner cross-sectional circle diameter of the lower end. 8. The liquid flow system of a flow cytometer according to claim 1, wherein the flushing pump (31) adopts a stepper motor to drive a syringe pump with a syringe structure, and has a volume of 600-1000 milliliters; Both the sample pump (28) and the microbead pump (29) are syringe pumps driven by a stepping motor with a volume of 500 to 600 microliters; the first auxiliary sheath pump (17), the second Both the auxiliary sheath fluid pump (18) and the main sheath fluid pump (30) are syringe pumps driven by a stepping motor with a volume of 30-40 milliliters. 9. The liquid flow system of a flow cytometer according to claim 1, wherein the air bubble detection module (24) is made of quartz glass or organic resin, is a cuboid structure or a cube structure, and has a square structure inside. channel, three interfaces are left on the outside to communicate with the internal channel; the ventilation module (26) is a cuboid structure or a cube structure, with a square channel inside, and a plurality of interfaces are left on the outside to communicate with the internal channel; the flushing buffer (35) is made of quartz Made of glass or organic resin, it is a cuboid or cube structure with a square channel inside and three interfaces on the outside to communicate with the inner channel. 10. The liquid flow system of a kind of flow cytometer according to claim 1, characterized in that, when the liquid flow system is in the low sample injection rate working mode, the first auxiliary sheath fluid pump (17) and the second The auxiliary sheath liquid pump (18) does not work, the first adjustment area inlet (9) and the second adjustment area inlet (10) are closed, and the main sheath liquid pump (30) passes through the sixth reversing valve (45) from the sheath liquid pool ( 50) to inhale the main sheath liquid, the main sheath liquid is pushed by the main sheath liquid pump (30) to enter the main sheath liquid inlet (6) and enter the inlet area (1), and the sample pump (28) passes through the fourth reversing valve (43) Inhale the sample solution from the sample injection tube (47), and push the sample solution from the sample flow tube (12) into the converging area (2) by the sample pump (28). If microbeads need to be detected, the microbead pump (29) passes through The third reversing valve (42) sucks the microbead liquid from the microbead sampling tube (48), and the microbead pump (29) pushes the microbead liquid to flow into the converging area (2) from the sample flow tube (12), and the main sheath Liquid, sample liquid and microbead liquid converge in the converging area (2) through the principle of hydrodynamic focusing and form a stable laminar flow, and the main sheath liquid wraps the sample liquid and microbead liquid and flows into the comprehensive adjustment area (11) and The detection area (4) flows into the waste liquid pool (49) from the auxiliary area (5) after detection; when the liquid flow system is in the high sample injection rate working mode, the inlet of the first adjustment area (9) and the inlet of the second adjustment area (10) conduction, the first auxiliary sheath liquid pump (17) and the second auxiliary sheath liquid pump (18) pass through the first reversing valve (15) and the second reversing valve (16) respectively from the sheath liquid pool (50 ), the auxiliary sheath liquid enters the comprehensive adjustment area (11) from the first adjustment area inlet (9) and the second adjustment area inlet (10) respectively, keeping the main sheath liquid flow rate and the sample liquid flow rate constant, Adjust the flow rate of the auxiliary sheath fluid so that it compresses the stable laminar flow flowing in from the converging area (2), the sample liquid is compressed and maintains a stable laminar flow, and the compressed stable laminar flow enters the detection area (4), and after detection, it flows from The auxiliary area (5) flows into the waste liquid pool (49), the flow rate of the main sheath liquid is greater than or equal to the flow rate of the auxiliary sheath liquid, and the flow rate of the auxiliary sheath liquid is greater than the sampling rate of the sample liquid.